Use of human plasma hyaluronidase in cancer treatment

An antibody that specifically binds to native human plasma hyaluronidase (hpHAse) produced by the process comprising the steps of:

• (a) incubating a candidate antibody with a sample comprising native hpHAse, said incubating being for a time sufficient for formation of antibody-hpHAse complexes;
• (b) contacting the sample with an insoluble support having anti-antibody and detectably-labeled hyaluronic acid bound thereto for a time sufficient for formation of anti-antibody-candidate antibody-hpHAse complexes;
• and
• (c) exposing the sample in contact with the support to an acidic pH of about 3.4 to 3.7, thereby allowing hpHAse in the antibody-hpHAse complex to degrade the detectably labeled hyaluronic acid;
• wherein samples associated with hyaluronic acid degradation comprise an anti-hpHAse antibody.

A method for screening candidate antibodies for binding to a native acid active hyaluronidase (aaHAse), the method comprising the steps of:

• (a) incubating a candidate antibody with a sample comprising native aaHAse, said incubating being for a time sufficient for formation of antibody-aaHAse complexes;
• (b) contacting the sample with an insoluble support having anti-antibody and detectably-labeled hyaluronic acid bound thereto for a time sufficient for formation of anti-antibody-candidate antibody-hpHAse complexes;
• and
• (c) exposing the sample in contact with the support to an acidic pH of about 3.4 to 3.7, thereby allowing hpHAse in the antibody-aaHAse complex to degrade the detectably labeled hyaluronic acid;
• wherein samples associated with hyaluronic acid degradation comprise an anti-aaHAse antibody.

A method for identifying a patient having or susceptible to a condition associated with a LuCa-1 defect, the method comprising the steps of:

• contacting a sample from the patient with an anti-hpHAse antibody, the sample being selected from the group consisting of tissue, blood, plasma, serum, and urine, said contacting being for a time sufficient for formation of anti-hpHAse antibody-hpHAse complexes;
• detecting the amount of hpHAse present in the sample;
• and
• comparing the amount of hpHAse detected in the sample with an amount of hpHAse in a control sample containing a known amount of hpHAse correlated with a normal level of hpHAse;
• wherein detection of an amount of hpHAse in the patient sample that is less than the amount of hpHAse in the control sample is indicative of a LuCa-1 defect in the patient.

A method of purifying a native acid active hyaluronidase (aaHAse) from a sample, the method comprising the steps of:

• (a) dissolving a sample suspected of containing an aaHAse in a solution at a temperature substantially less than room temperature, the solution comprising a non-ionic detergent;
• (b) raising the temperature of the solution to a temperature substantially greater than room temperature, said raising resulting in the formation of a detergent-rich phase comprising aaHAse and a detergent-poor phase;
• and
• (c) isolating aaHAse from the detergent-rich phase.

A method of making a recombinat human plasma hylauronidase polypeptide, the method comprising the steps of:

• introducing into a host cell a nucleotide sequence encoding a human plasma hylaruonidase polypeptide;
• culturing the host cell so as to allow for expression of the polypeptide from the introduced sequence;
• and
• isolating human plasma hylauronidase polypeptide.